CN108329330B

CN108329330B - 2-benzyloxy phenyl oxazolopyridine compound and pharmaceutical application thereof

Info

Publication number: CN108329330B
Application number: CN201710042835.7A
Authority: CN
Inventors: 叶德泳; 齐翔宇; 李亚莉; 莫明广; 周璐; 楚勇; 王鹏辉
Original assignee: Fudan University
Current assignee: Fudan University
Priority date: 2017-01-20
Filing date: 2017-01-20
Publication date: 2021-05-04
Anticipated expiration: 2037-01-20
Also published as: CN108329330A

Abstract

The invention belongs to the field of pharmaceutical chemistry, and relates to a 2-benzyloxy phenyl oxazolopyridine compound shown in a formula I structure and a pharmaceutical application thereof, wherein R, X, Y and Z are consistent with detailed description in the invention content. The compounds can inhibit the activity of sphingomyelin synthase and can be used for preparing small molecule inhibitors of sphingomyelin synthase. The invention provides aA compound shown as a formula I structure or a pharmaceutical composition taking the compound as an effective active ingredient and application thereof in preparing medicaments for preventing and treating diseases caused by abnormal increase of sphingomyelin level; the diseases caused by abnormal increase of sphingomyelin level comprise metabolic syndromes such as atherosclerosis, fatty liver, obesity and type II diabetes.

Description

2-benzyloxy phenyl oxazolopyridine compound and pharmaceutical application thereof

Technical Field

The invention belongs to the field of medicinal chemistry, and relates to a 2-benzyloxy phenyl oxazolopyridine compound and a medicinal application thereof, in particular to a 2-benzyloxy phenyl oxazolopyridine compound and an application thereof in preparation of a sphingomyelin synthase inhibitor.

Background

Data show that, in China, with the development of economic society and the aging of population, the incidence and mortality of cardiovascular and cerebrovascular diseases are remarkably increased to 2 nd of the total causes of death except tumors, and the incidence and mortality become one of the main diseases harmful to human health. According to the prediction, the number of cardiovascular disease patients in China will be increased by 2130 ten thousand and the number of cardiovascular disease deaths will be increased by 770 ten thousand in 2010-2030 years. Research shows that Atherosclerosis (AS) is a main cause of diseases such AS cardiovascular and cerebrovascular diseases, so that the prevention of AS is a key link for preventing cardiovascular and cerebrovascular diseases, and research of anti-atherosclerosis drugs becomes a hot spot in the current drug research and development field.

Research reports disclose that the main pathological manifestations of atherosclerosis are lipid deposition, mononuclear and lymphocyte infiltration of the affected intima of arteries and proliferation, migration of vascular smooth muscle cells, foam cell and fibrous plaque formation, which in turn causes vessel wall sclerosis, luminal narrowing and thrombosis. Although the molecular pathology has not been fully elucidated, it is recognized that among the known factors, dyslipidemia is the most important inducer of atherosclerotic formation, and abnormal expression of lipid components is closely related to the formation of atherosclerotic and sclerotic plaques. In general, dyslipidemia refers to abnormal fat metabolism or transport, which results in higher than normal plasma lipids and increased blood viscosity, and is mainly manifested by increased low-density lipoprotein (LDL) and very low-density lipoprotein (VLDL) levels and decreased high-density lipoprotein (HDL) levels, so that lowering LDL and/or increasing HDL can play a role in regulating blood lipids, and the lipid regulating drug also becomes a main drug clinically used for anti-atherosclerosis. The currently clinically used lipid regulators mainly include statins, fibrates, bile acid binding resins, nicotinic acid and the like.

With the progress of research, various potential drug targets for resisting atherosclerosis are proposed: sphingomyelin synthase inhibitors, PPAR agonists, Cholesteryl Ester Transfer Protein (CETP) inhibitors, infusion apolipoproteins, liver X receptor activators, phospholipid transfer protein (PLTP) inhibitors, and the like; in particular, Sphingomyelin (SM) and its metabolic enzymes mediate a series of cellular processes while causing lipoprotein changes, suggesting that it plays an important role in the development of atherosclerosis.

Studies have shown that sphingomyelin can induce atherosclerosis through a variety of pathways: (1) inhibition of lipolysis of Triglycerides (TG) (Park TS, Panek RL, et al.Atherosclerosis. 2006, 189(2)264-72.); (2) delay the clearance of lipoprotein remnant leading to AS (Schlitt A, Hojjati MR, et al.J. Lipid Res. 2005, 46 (2)196 ℃ and 200 ℃); (3) affecting HDL-mediated reverse cholesterol transport, resulting in cholesterol-clearing disorders (Sano O, Kobayashi a, et al.J. Lipid Res. 2007, 48(11): 2377-84; Marmillot P, Patel S, et al. Metabolism. 2007, 56(2)251-9.); (4) related products of ceramide and SM synthesis or breakdown are regulators of cell proliferation, activation, apoptosis, affecting the growth and stabilization of atherosclerotic plaques (Park, t. -s.; Panek, r. l.; et al.Circulation. 2004, 1103465-; (5) SM-rich LDL has strong coagulationAggregation and adhesion, which results in macrophages more likely to accumulate in the arterial wall to form foam cells and thus promote AS (Fan Y, Shi S, et al.Arterioscler Thromb Vasc Biol. 2010, 30: 2114-20.)。

Epidemiological investigations have shown that there is an independent correlation between human SM levels and Atherosclerosis (AS), and that plasma SM concentration is an independent risk factor for atherosclerosis and has an indicative meaning in assessing the progression of atherosclerosis (Jiang, X. -C.; Paultre, F.; et al.Arterioscler Thromb Vasc Biol. 2000, 20: 2614-2618; Zhiqiang Li; Maria J. Basterr; et al. Biochimica et Biophysica Acta. 2005, 1735130-; animal experimental studies have shown that inhibition of de novo SM is indeed effective in reducing plasma cholesterol and triglyceride levels in apoE-KO mice, raising HDL-cholesterol levels, and thus preventing the development of AS lesions (Park, t. -s.; Panek, r. l.; et al).Circulation. 2004, 1103465-; thus, lowering plasma sphingomyelin levels or inhibiting SM synthesis can serve the purpose of slowing or preventing the development of atherosclerosis.

It has also been found that sphingomyelin synthase (SMS) regulates the synthesis of SM by ceramide (ceramide) and lecithin (PC), and is a key enzyme in the last step of the sphingomyelin de novo synthesis pathway. Further studies have found that SMS directly regulates SM levels, and that over-expression of SMS is a common phenomenon in atherosclerotic lesion tissues and is one of the key indicators of atherosclerotic lesion development (Xiaoan-cheng Jiang; Furcy Paultre; et al.Arterioscler Thromb Vasc Biol.2000, 20: 2614-2618；Zhiqiang Li; Tiruneh K. et al. Bioch Bioph Acta. 2007, 1771: 1186-1194.). There are three subtypes of SMS, namely SMS1 located in the trans-Golgi complex, SMS2 located in the cytoplasmic membrane, and SMS-related protein (SMSr), which has no SMS activity. In knockout experiments, lack of SMS1 and SMS2 alone resulted in a decrease in plasma sphingomyelin content by about 50%, and increased plasma ceramide levels in SMS2 knockout mice, whereas knockout of the SMS1 gene had no effect on this concentration (Jiang XC.Chin J Hypertens. 2014, 22(7)603-; the experimental results suggest that SMS1 is mainly distributed in macrophages, and SMS2 is mainly distributed in liver cells; animal experiments showed that SMS2 deficiency increased plasma apoE levels, whereas SMS2 overexpression decreased plasma apoE levels, contributing to AS formation (Liu J, Jiang XC. et al.Arterioscler Thromb Vasc Biol. 2009, 29850-856), the aortic arch atherosclerotic plaques of the SMS2 and apoE double gene knockout mouse model were significantly reduced, the lipid levels of SM, etc. in the brachiocephalic artery were significantly reduced, and no effect on the normal physiology of the mice was observed (Fan Y, Shi S, et al.Arterioscler Thromb Vasc Biol. 2010, 302114. 2120.), which shows that the reaction of synthesizing SM catalyzed by SMS is in the last stage of the sphingomyelin biosynthesis cycle, and the potential toxic and side effects caused by inhibiting the activity of SM are less; in conclusion, the research results show that the method for reducing the level of the sphingomyelin by inhibiting the sphingomyelin synthase can become a novel method for treating atherosclerosis, the sphingomyelin synthase has potential superiority as a novel target for resisting the atherosclerosis, and the sphingomyelin synthase inhibitor can become a novel therapeutic drug for resisting the atherosclerosis.

In addition, SMS2 deficiency was found to prevent high fat diet-induced obesity and insulin resistance, while large mature fatty plaques were difficult to observe in the liver of SMS2 knockout mice, suggesting that SMS2 is involved in the formation of fatty plaques in the liver and may induce the development of obesity and type ii diabetes (Susumu Mitsutake, Kota Zama, et al.J Biol Chem. 2011, 286(32): 28544-28555.). Reduction of SM in plasma due to SMS2 deficiency can improve insulin sensitivity in animal tissues and throughout the body (Li Z, Zhang H, et al.Mol Cell Biol. 2011, 31(20)4205 and 4218.) therefore, the small molecule inhibitors of sphingomyelin synthase can prevent and treat metabolic syndromes such as obesity, fatty liver and type II diabetes.

One of the sphingomyelin synthase inhibitors reported in the literature is D609(Aimin Meng; Chiara Luberto; et al.Exper Cell Res. 2004, 292385-392.), the enzyme inhibitory activity of which was weak (SMS inhibitory activity IC)₅₀= 402.7 ± 8.6 μ M, mice livers were homogenized; SMS2 inhibitory Activity IC₅₀= 224.6 ± 4.5 μ M, high expression insect cell homogenate for SMS2), and contains a orthosulfonate in the chemical structure making the structure highly unstable (Bai, a. et al.J Pharmacol Exp Ther. 2004, 3091051-1059.), short half-life; in addition, the method of homologous modeling is researched and constructed for the first timehThree-dimensional protein structure model of SMS1 (human SMS1 type) (Zhang Ya; Lin Fu; et al.Chin J Chem 2011, 292421-2429.) Using this three-dimensional protein structural model and the active sites for validated enzyme-substrate binding, compound D2 (Xiaoodong Deng, Fu Lin, et al) was discovered as a small molecule inhibitor of sphingomyelin synthase.Eur J Med Chem,2014, 731-7.) the inhibitory activity against SMS in vitro (SMS inhibitory activity IC)₅₀Homogenizing mouse liver at 23.5 + -3.2 μ M; SMS2 inhibitory Activity IC₅₀= 13.3 ± 0.26 μ M, for insect cells highly expressed by SMS2) is higher than D609, but the following defects still exist: the inhibitory activity on SMS2 is to be improved, and the inhibitor contains cyano groups with high potential toxicity risk and has poor water solubility, stability and other physicochemical properties.

Based on the current situation of the prior art, the inventor intends to provide 2-benzyloxyphenyl oxazolopyridine compounds and pharmaceutical uses thereof, and particularly relates to 2-benzyloxyphenyl oxazolopyridine compounds and uses thereof in preparation of sphingomyelin synthase inhibitors.

Disclosure of Invention

The invention aims to overcome the defects and shortcomings of the prior art and provides a 2-benzyloxyphenyl oxazolopyridine compound and a pharmaceutical application thereof, in particular to a 2-benzyloxyphenyl oxazolopyridine compound and an application thereof in preparation of a sphingomyelin synthase inhibitor and an application thereof in preparation of a medicament for preventing or treating atherosclerosis, fatty liver, obesity, type II diabetes and related metabolic syndrome.

The first object of the invention is to provide 2-benzyloxy phenyl oxazole pyridine compound; the 2-benzyloxy phenyl oxazolopyridine compound is free alkali or salt with a structure shown in a formula I,

（I）

wherein X, Y and Z are selected from carbon atoms or nitrogen atoms, and can be carbon atoms, and at most only one is nitrogen atom; r is selected from any one or two of H, halogen, nitro, cyano, alkyl, alkoxy and trifluoromethoxy; the phenyl group in which R is substituted by pyridyl; the substituted benzyloxy group may be in the ortho-or para-position.

Further, the substituent R may beo-F，m-F，p-F，o-Cl，m-Cl，p-Cl，o-Br，m-Br，p-Br，o-NO₂，m-NO₂，p-NO₂，o-CN，m-CN，p-CN，o-OCF₃，m-OCF₃，p-OCF₃，o-CH₃，m-CH₃，p-CH₃，o-OCH₃，m-OCH₃，p-OCH₃Either one or both.

Further described are compounds of formula I-1 through formula I-33:


a general formula I-1,	A general formula I-2,	A general formula I-3,

A formula I-4,	A formula I-5,	A formula I-6,

A formula I-7,	A formula I-8,	A formula I-9,

A formula I-10,	A formula I-11,	A formula I-12,

A formula I-13,	A formula I-14,	A formula I-15,

A formula I-16,	A formula I-17,	A formula I-18,

A general formula I-19,	A formula I-20,	A formula I-21,

A formula I-22,	A formula I-23,	A formula I-24,

A formula I-25,	A formula I-26,	A formula I-27,

A formula I-28,	A formula I-29,	A formula I-30,

A formula I-31,	A formula I-32,	Formula I-33.

The solvate of the 2-benzyloxy phenyl oxazole pyridine compound of the invention also belongs to the protection scope of the invention, and the solvent is preferably water, ethanol or methanol.

The second purpose of the invention is to provide the application of the 2-benzyloxy phenyl oxazolopyridine compound shown in the formula I in the preparation of small molecule inhibitors of sphingomyelin synthase. The invention adopts a High Performance Liquid Chromatography (HPLC) fluorescence quantitative detection method reported in the literature to measure the inhibition activity of 2-benzyloxy phenyl oxazolopyridine compounds shown in formula I on sphingomyelin synthase (Xiaoodong Deng; Hong Sun; et al.Analytical Letters, 2012, 45(12)1581. 1589.) the change in the activity of the inhibitor on the conversion of ceramide to sphingomyelin by sphingomyelin synthase was calculated from the change in the amounts of NBD-ceramide and NBD-sphingomyelin contained therein.

An activity test experiment based on a High Performance Liquid Chromatography (HPLC) fluorescence quantitative method shows that the 2-benzyloxy phenyl oxazolopyridine compound shown in the formula I has sub-micromolar sphingomyelin synthase inhibition activity and is an effective component for inhibiting sphingomyelin synthase; and the selectivity of the SMS2 subtype is higher in the two subtypes of SMS1 and SMS 2. The High Performance Liquid Chromatography (HPLC) fluorescence quantitative method is adopted to detect the inhibition activity of the compound on sphingomyelin synthase 2(SMS2) as follows:

1) the inhibition rate of (2- (benzyloxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-1) at 10. mu.M is 69%;

2) the inhibition rate of (2- ((2-chlorobenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-2) at 10 mu M is 52%;

3) the inhibition rate of (2- ((3-chlorobenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-3) at 10 mu M is 74%;

4) the inhibition rate of (2- ((4-chlorobenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-4) at 10 mu M is 20%;

5) the inhibition rate of (2- ((4-methylbenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-5) at 10 mu M is 22%;

6) the inhibition rate of (2- ((2-fluorobenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-6) at 10 mu M is 46%;

7) the inhibition rate of (2- ((3-fluorobenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-7) at 10 mu M is 80%;

8) the inhibition rate of (2- ((4-fluorobenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-8) at 10 mu M is 30%;

9) the inhibition rate of (2- ((2-methylbenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-9) at 10 mu M is 71%;

10) the inhibition rate of (2- ((3-methylbenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-10) at 10 mu M is 57%;

11) the inhibition rate of (2- ((2-nitrobenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-11) at 10 mu M is 32%;

12) the inhibition rate of (2- ((3-nitrobenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-12) at 10 mu M is 65%;

13) the inhibition rate of (2- ((4-nitrobenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-13) at 10 mu M is 10%;

14) the inhibition rate of (2- ((2-methoxybenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-14) at 10 mu M is 54%;

15) the inhibition rate of (2- ((3-methoxybenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-15) at 10 mu M is 93%;

16) the inhibition rate of (2- ((4-methoxybenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-16) at 10 mu M is 9%;

17) the inhibition rate of (2- (2-ethylbenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-17) at 10 mu M is 78%;

18) the inhibition rate of (2- ((2- (trifluoromethoxy) benzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-18) at 10 μ M is 57%;

19) the inhibition rate of (2- ((3- (trifluoromethoxy) benzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-19) at 10 mu M is 28%;

20) the inhibition rate of (2- ((4- (trifluoromethoxy) benzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-20) at 10 mu M is 14%;

21) the inhibition rate of 2- ((2- (oxazolo [5,4-b ] pyridin-2-yl) phenoxy) methyl) benzonitrile (formula I-21) at 10. mu.M was 31%;

22) the inhibition of 3- ((2- (oxazolo [5,4-b ] pyridin-2-yl) phenoxy) methyl) benzonitrile (formula i-22) at 10 μ M was 61%;

23) the inhibition of 4- ((2- (oxazolo [5,4-b ] pyridin-2-yl) phenoxy) methyl) benzonitrile (formula i-23) at 10 μ M was 14%;

24) the inhibition rate of (2- ((3-fluoro-5-methoxybenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-24) at 10 mu M is 65%;

25) the inhibition rate of (2- ((3, 5-difluorobenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-25) at 10 mu M is 62%;

26) the inhibition rate of (2- ((3-fluoro-2-methylbenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-26) at 10 mu M is 89%;

27) the inhibition rate of (2- ((5-fluoro-2-methylbenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-27) at 10 mu M is 66%;

28) the inhibition rate of (2- ((3-methoxy-2-methylbenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-28) at 10 mu M is 24%;

29) the inhibition rate of (2- ((5-methoxy-2-methylbenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-29) at 10 mu M is 64%;

30) the inhibition rate of (2- (pyridine-2-yl methoxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-30) at 10 mu M is 8%;

31) the inhibition rate of (2- ((2, 6-dimethylbenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-31) at 10 mu M is 83%;

32) the inhibition of (2- (benzyloxy) phenyl) benzo [ d ] oxazole (formula I-32) at 10. mu.M was 25%;

33) the inhibition of (4- (benzyloxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-33) at 10. mu.M was 8%.

The invention further aims to provide application of the 2-benzyloxy phenyl oxazolopyridine compound shown in the formula I and the solvate thereof in preparing medicines for preventing and treating diseases caused by abnormal increase of sphingomyelin level, such as atherosclerosis, fatty liver, obesity, diabetes type II and other related metabolic syndromes.

The above medicine may further comprise one or more pharmaceutically acceptable carriers, which include conventional diluents, excipients, fillers, binders, wetting agents, disintegrants, absorption enhancers, surfactants, adsorption carriers, lubricants, etc. in the pharmaceutical field, and if necessary, flavoring agents, sweeteners, etc.

The 2-benzyloxy phenyl oxazolopyridine compounds provided by the invention are sphingomyelin synthase inhibitors with novel structures, have subtype selectivity and sub-micromolar molecular level inhibition activity, have good potential and application prospect, and can be further prepared into medicines for treating atherosclerosis, fatty liver, obesity, type II diabetes and other related metabolic syndromes.

Detailed Description

Example 1: preparation of (2- (benzyloxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-1)

Synthesis of (oxazolo [5,4-b ] pyridin-2-yl) phenol (Compound 1 c)

1.0 g (9.08 mmol, 1.05 eq) of 3-amino-2-hydroxypyridine was dissolved in 34.6 mL of an aqueous solution of sodium hydroxide (1 mol/L, 34.60 mmol, 4.0 eq) at room temperature, and after stirring the mixture uniformly, 1.4 g (8.65 mmol, 1.0 eq) of 2-trifluoromethylphenol, 80^oC, heating the mixture in oil bath for 2.0 h. TLC detection raw material disappeared, stop heating. Cooling to room temperature, extracting with dichloromethane, combining organic phases, washing with a saturated sodium chloride solution, drying with anhydrous sodium sulfate, purifying by PE-EA system column chromatography, and concentrating to obtain a compound 1c white solid 1.40 g with a yield of 76.3%;

through detection, the structure is correct, and the detection result is as follows:

compound 1c m.p.153.7-154.5^oC; ¹H NMR (400 MHz, DMSO) δ 11.02 (s, 1H), 8.37 (dd, J = 4.9, 1.2 Hz, 1H), 8.26 (dd, J = 7.8, 1.2 Hz, 1H), 7.99 (dd, J = 7.8, 1.3 Hz, 1H), 7.51 (dd, J = 7.7, 5.0 Hz, 2H), 7.12 – 7.02 (m, 2H); MS(ESI) (m/z): 213.1(M+H)⁺.

Synthesis of di- (2- (benzyloxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-1)

310 mg (1.46 mmol, 1.0 eq) of (oxazolo [5,4-b ] pyridin-2-yl) phenol (compound 1 c) and 404 mg (2.92 mmol, 2.0 eq) of anhydrous potassium carbonate were mixed and dissolved in 15 ml of acetonitrile, stirred uniformly, 300 mg of benzyl bromide (1.75 mmol, 1.2 eq) was slowly added at room temperature, reacted at room temperature overnight, and the starting material was lost by TLC. Evaporating to remove acetonitrile, extracting with water and DCM to obtain liquid, washing water phase with DCM, mixing organic phases, washing twice with saturated sodium chloride solution, adding anhydrous sodium sulfate, stirring, drying, and purifying by PE-EA system column chromatography to obtain white loose solid I-1 with yield of 80%;

formula I-1 m.p.91.5-92.7^oC; ¹H NMR (400 MHz, DMSO) δ 8.37 (dd, J = 4.9, 1.5 Hz, 1H), 8.27 (dd, J = 7.8, 1.5 Hz, 1H), 8.08 (dd, J = 7.8, 1.7 Hz, 1H), 7.63 – 7.56 (m, 2H), 7.49 (dd, J = 7.8, 4.9 Hz, 1H), 7.41 – 7.27 (m, 4H), 7.16 (t, J = 7.5 Hz, 1H), 5.35 (s, 2H); MS(ESI) (m/z): 303.1(M+H)⁺.。

Example 2: synthesis of Compounds of formulae I-2, I-3, I-4, I-5, I-6, I-7, I-8, I-9, I-10, I-11, I-12, I-13, I-17, I-18, I-19, I-20, I-21, I-22, I-23, I-24, I-25

Formula I	R
		Ⅰ-2	2-Cl
Ⅰ-3	3-Cl
		Ⅰ-4	4-Cl
Ⅰ-5	4-CH₃
		Ⅰ-6	2-F
Ⅰ-7	3-F
		Ⅰ-8	4-F
Ⅰ-9	2-CH₃
		Ⅰ-10	3-CH₃
Ⅰ-11	2-NO₂
		Ⅰ-12	3-NO₂
Ⅰ-13	4-NO₂
		Ⅰ-17	2-Et
Ⅰ-18	2-OCF₃
		Ⅰ-19	3-OCF₃
Ⅰ-20	4-OCF₃
		Ⅰ-21	2-CN
Ⅰ-22	3-CN
		Ⅰ-23	4-CN
Ⅰ-24	3-F; 5-OCH3
		Ⅰ-25	3, 5-difluoro

Referring to the conditions for the second synthesis of compound of formula I-1 in example 1, compounds of formulae I-2 to I-13 and formulae I-17 to I-25 were obtained from (oxazolo [5,4-b ] pyridin-2-yl) phenol (compound 1 c) and the corresponding commercially available substituted benzyl bromides (compound 2d to 13d and compound 17d to 25 d): (2- ((2-chlorobenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-2); (2- ((3-chlorobenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-3); (2- ((4-chlorobenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-4); (2- ((4-methylbenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-5); (2- ((2-fluorobenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-6); (2- ((3-fluorobenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-7); (2- ((4-fluorobenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-8); (2- ((2-methylbenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-9); (2- ((3-methylbenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-10); (2- ((2-nitrobenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-11); (2- ((3-nitrobenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-12); (2- ((4-nitrobenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-13); (2- (2-ethylbenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-17); (2- ((2- (trifluoromethoxy) benzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-18); 2- ((3- (trifluoromethoxy) benzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-19); (2- ((4- (trifluoromethoxy) benzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-20); 2- ((2- (oxazolo [5,4-b ] pyridin-2-yl) phenoxy) methyl) benzonitrile (formula i-21); 3- ((2- (oxazolo [5,4-b ] pyridin-2-yl) phenoxy) methyl) benzonitrile (formula i-22); 4- ((2- (oxazolo [5,4-b ] pyridin-2-yl) phenoxy) methyl) benzonitrile (formula i-23); (2- ((3-fluoro-5-methoxybenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-24); (2- ((3, 5-difluorobenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-25);

formula I-2 m.p.119.9-121.3^oC; ¹H NMR (400 MHz, DMSO) δ 8.37 (dd, J = 4.9, 1.5 Hz, 1H), 8.28 (dd, J = 7.8, 1.5 Hz, 1H), 8.12 (dd, J = 7.8, 1.6 Hz, 1H), 7.99 (dd, J = 7.4 Hz, 1H), 7.68 – 7.58 (m, 1H), 7.49 (dd, J = 7.8, 5.0 Hz, 2H), 7.44 – 7.32 (m, 3H), 7.20 (t, J = 7.6 Hz, 1H), 5.37 (s, 2H). MS(ESI) (m/ z): 337.1(M+H)⁺.

Formula I-3 m.p.130.6-132.4^oC; ¹H NMR (400 MHz, DMSO) δ 8.39 (dd, J = 4.9, 1.5 Hz, 1H), 8.27 (dd, J = 7.8, 1.5 Hz, 1H), 8.11 (dd, J = 7.8, 1.6 Hz, 1H), 7.84 (s, 1H), 7.66 – 7.58 (m, 1H), 7.56 – 7.47 (m, 2H), 7.39 (dt, J = 26.5, 8.1 Hz, 3H), 7.19 (t, J = 7.6 Hz, 1H), 5.37 (s, 2H). MS(ESI) (m/z): 337.1(M+H)⁺.

Formula I-4 m.p.124.8-125.8^oC; ¹H NMR (400 MHz, DMSO) δ 8.37 (dd, J = 4.9, 1.5 Hz, 1H), 8.29 (dd, J = 7.8, 1.5 Hz, 1H), 8.09 (dd, J = 7.8, 1.7 Hz, 1H), 7.66 – 7.57 (m, 3H), 7.51 – 7.42 (m, 3H), 7.33 (d, J = 8.4 Hz, 1H), 7.17 (t, J = 7.6 Hz, 1H), 5.34 (s, 2H). MS(ESI) (m/z): 337.1(M+H)⁺.

Formula I-5 m.p.91.2-91.5^oC; ¹H NMR (400 MHz, DMSO) δ 8.37 (dd, J = 4.9, 1.5 Hz, 1H), 8.27 (dd, J = 7.8, 1.6 Hz, 1H), 8.07 (dd, J = 7.8, 1.7 Hz, 1H), 7.59 (ddd, J = 8.6, 7.4, 1.8 Hz, 1H), 7.51 – 7.43 (m, 3H), 7.33 (d, J = 8.4 Hz, 1H), 7.20 – 7.12 (m, 3H), 5.29 (s, 2H), 2.27 (s, 3H). MS(ESI) (m/z): 317.2(M+H)⁺.

Formula I-6 m.p.108.6-109.5^oC; ¹H NMR (400 MHz, DMSO) δ 8.36 (dd, J = 4.9, 1.3 Hz, 1H), 8.26 (dd, J = 7.8, 1.1 Hz, 1H), 8.09 (dd, J = 7.8, 1.5 Hz, 1H), 7.84 (dd, J = 10.9, 4.6 Hz, 1H), 7.66 – 7.59 (m, 1H), 7.50 – 7.45 (m, 1H), 7.38 (dd, J = 15.4, 7.4 Hz, 2H), 7.22 (ddd, J = 22.6, 10.2, 6.0 Hz, 3H), 5.37 (s, 3H). MS(ESI) (m/z): 321.0(M+H)⁺.

Formula I-7 m.p.111.5-113.6^oC; ¹H NMR (400 MHz, DMSO) δ 8.38 (d, J = 4.9 Hz, 1H), 8.25 (d, J = 7.8 Hz, 1H), 8.10 (d, J = 7.7 Hz, 1H), 7.62 (t, J = 7.9 Hz, 1H), 7.54 – 7.47 (m, 2H), 7.46 – 7.38 (m, 2H), 7.34 (d, J = 8.5 Hz, 1H), 7.15 (dt, J = 16.3, 7.1 Hz, 2H), 5.37 (s, 2H). MS(ESI) (m/z): 321.0(M+H)⁺.

Formula I-8 m.p.95.8-97.0^oC; ¹H NMR (400 MHz, DMSO) δ 8.37 (dd, J = 4.9, 1.6 Hz, 1H), 8.28 (dd, J = 7.8, 1.5 Hz, 1H), 8.09 (dd, J = 7.8, 1.7 Hz, 1H), 7.68 – 7.58 (m, 3H), 7.49 (dd, J = 7.8, 4.9 Hz, 1H), 7.35 (d, J = 8.2 Hz, 1H), 7.26 – 7.13 (m, 3H), 5.33 (s, 2H). MS(ESI) (m/z): 321.0(M+H)⁺.

Formula I-9 m.p.100.3-102.4^oC; ¹H NMR (400 MHz, DMSO) δ 8.36 (dd, J = 4.9, 1.5 Hz, 1H), 8.24 (dd, J = 7.8, 1.6 Hz, 1H), 8.08 (dd, J = 7.8, 1.7 Hz, 1H), 7.69 – 7.59 (m, 2H), 7.50 – 7.40 (m, 2H), 7.22 – 7.13 (m, 4H), 5.30 (s, 2H), 2.36 (s, 3H). MS(ESI) (m/z): 317.2(M+H)⁺.

Formula I-10 m.p.79.0-82.0^oC; ¹H NMR (400 MHz, DMSO) δ 8.38 (dd, J = 4.9, 1.5 Hz, 1H), 8.27 (dd, J = 7.8, 1.5 Hz, 1H), 8.08 (dd, J = 7.8, 1.6 Hz, 1H), 7.64 – 7.57 (m, 1H), 7.52 – 7.46 (m, 2H), 7.36 – 7.31 (m, 2H), 7.25 (t, J = 7.6 Hz, 1H), 7.16 (t, J = 7.5 Hz, 1H), 7.10 (d, J = 7.5 Hz, 1H), 5.30 (s, 2H), 2.31 (s, 3H). MS(ESI) (m/z): 317.2(M+H)⁺.

Formula I-11 m.p.152.0-153.2^oC; ¹H NMR (400 MHz, DMSO) δ 8.40 – 8.28 (m, 3H), 8.18 – 8.12 (m, 2H), 7.87 (t, J = 7.5 Hz, 1H), 7.67 – 7.59 (m, 2H), 7.51 (dd, J = 7.8, 5.0 Hz, 1H), 7.37 (d, J = 8.5 Hz, 1H), 7.21 (t, J = 7.5 Hz, 1H), 5.69 (s, 2H). MS(ESI) (m/z): 348.1(M+H)⁺.

Formula I-12 m.p.180.0-183.2^oC; ¹H NMR (400 MHz, DMSO) δ 8.80 (s, 1H), 8.38 (dd, J = 4.9, 1.4 Hz, 1H), 8.29 (dd, J = 7.8, 1.3 Hz, 1H), 8.19 – 8.11 (m, 2H), 8.00 (d, J = 7.7 Hz, 1H), 7.66 (ddd, J = 15.7, 12.3, 4.7 Hz, 2H), 7.52 (dd, J = 7.8, 4.9 Hz, 1H), 7.37 (d, J = 8.4 Hz, 1H), 7.20 (t, J = 7.5 Hz, 1H), 5.48 (s, 2H). MS(ESI) (m/z): 348.1(M+H)⁺.

Formula I-13 m.p.170.5-173.3^oC; ¹H NMR (400 MHz, DMSO) δ 8.39 (dd, J = 4.9, 1.5 Hz, 1H), 8.35 (dd, J = 7.8, 1.5 Hz, 1H), 8.29 (d, J = 8.8 Hz, 2H), 8.14 (dd, J = 7.8, 1.7 Hz, 1H), 7.94 (d, J = 8.9 Hz, 2H), 7.65 – 7.61 (m, 1H), 7.51 (dd, J = 7.8, 4.9 Hz, 1H), 7.35 (d, J = 8.1 Hz, 1H), 7.22 – 7.18 (m, 1H), 5.52 (s, 2H). MS(ESI) (m/z): 348.1(M+H)⁺.

Formula I-17¹H NMR (400 MHz, DMSO-d ₆) δ 8.35 (dd, J = 5.0, 1.6 Hz, 1H), 8.24 (dd, J = 7.8, 1.6 Hz, 1H), 8.07 (dd, J = 7.7, 1.8 Hz, 1H), 7.71 – 7.58 (m, 2H), 7.51 – 7.40 (m, 2H), 7.31 – 7.12 (m, 4H), 5.32 (s, 2H), 2.71 (q, J = 7.5 Hz, 2H), 1.12 (t, J = 7.5 Hz, 3H). MS(ESI) (m/z): 330.1(M+H)⁺.

Formula I-18¹H NMR (400 MHz, DMSO-d ₆) δ 8.29 (dd, J = 5.1, 1.5 Hz, 1H), 8.17 (dd, J = 7.8, 1.6 Hz, 1H), 8.00 (dd, J = 7.9, 1.7 Hz, 1H), 7.83 (d, J = 7.6 Hz, 1H), 7.58 (t, J = 7.5 Hz, 1H), 7.47 – 7.28 (m, 5H), 7.15 (t, J = 7.6 Hz, 1H), 5.28 (s, 2H). MS(ESI) (m/z): 386.1(M+H)⁺.

Formula I-19¹H NMR (400 MHz, DMSO-d ₆) δ 8.32 (dd, J = 5.0, 1.5 Hz, 1H), 8.15 (dd, J = 7.9, 1.6 Hz, 1H), 8.03 (dd, J = 8.0, 1.7 Hz, 1H), 7.63 (s, 1H), 7.58 (t, J = 7.7 Hz, 1H), 7.54 – 7.44 (m, 3H), 7.31 (d, J = 8.5 Hz, 1H), 7.23 (d, J = 8.1 Hz, 1H), 7.16 (t, J = 7.6 Hz, 1H), 5.33 (s, 2H). MS(ESI) (m/z): 386.1(M+H)⁺.

Formula I-20¹H NMR (400 MHz, DMSO-d ₆) δ 8.38 (dd, J = 5.0, 1.6 Hz, 1H), 8.30 (dd, J = 7.8, 1.6 Hz, 1H), 8.10 (dd, J = 7.8, 1.7 Hz, 1H), 7.78 – 7.71 (m, 2H), 7.62 (ddd, J = 8.9, 7.4, 1.8 Hz, 1H), 7.49 (dd, J = 7.8, 4.9 Hz, 1H), 7.37 (dd, J = 16.9, 8.3 Hz, 3H), 7.22 – 7.13 (m, 1H), 5.39 (s, 2H). MS(ESI) (m/z): 386.1(M+H)⁺.

Formula I-21¹H NMR (400 MHz, DMSO-d ₆) δ 8.35 (dd, J = 5.1, 1.5 Hz, 1H), 8.25 (dd, J = 7.7, 1.5 Hz, 1H), 8.12 (dd, J = 8.0, 1.7 Hz, 1H), 8.01 (d, J = 7.8 Hz, 1H), 7.92 – 7.84 (m, 1H), 7.82 – 7.72 (m, 1H), 7.65 (ddd, J = 9.1, 7.3, 1.8 Hz, 1H), 7.59 – 7.51 (m, 1H), 7.47 (dd, J = 7.9, 4.9 Hz, 1H), 7.40 (d, J = 8.4 Hz, 1H), 7.22 (t, J = 7.6 Hz, 1H), 5.48 (s, 2H). MS(ESI) (m/z): 327.1(M+H)⁺.

Formula I-22¹H NMR (400 MHz, DMSO-d ₆) δ 8.41 – 8.34 (m, 1H), 8.31 – 8.21 (m, 2H), 8.16 – 8.08 (m, 1H), 7.91 (d, J = 7.8 Hz, 1H), 7.78 (d, J = 7.6 Hz, 1H), 7.62 (q, J = 7.8, 7.3 Hz, 2H), 7.50 (dd, J = 7.9, 4.9 Hz, 1H), 7.34 (d, J = 8.3 Hz, 1H), 7.19 (t, J = 7.6 Hz, 1H), 5.40 (s, 2H). MS(ESI) (m/z): 327.1(M+H)⁺.

Formula I-23¹H NMR (400 MHz, DMSO-d ₆) δ 8.38 (dd, J = 5.0, 1.6 Hz, 1H), 8.30 (dd, J = 7.8, 1.6 Hz, 1H), 8.12 (dd, J = 7.7, 1.8 Hz, 1H), 7.92 – 7.80 (m, 4H), 7.61 (ddd, J = 8.8, 7.5, 1.9 Hz, 1H), 7.54 – 7.46 (m, 1H), 7.33 (d, J = 8.5 Hz, 1H), 7.28 – 7.14 (m, 1H), 5.45 (s, 2H). MS(ESI) (m/z): 327.1(M+H)⁺.

Formula I-24¹H NMR (300 MHz, DMSO) δ 8.40 (d, J = 4.8 Hz, 1H), 8.26 (d, J= 7.8 Hz, 1H), 8.12 (d, J = 7.7 Hz, 1H), 7.64 (t, J = 7.9 Hz, 1H), 7.52 (dd, J = 7.7, 5.1 Hz, 1H), 7.35 (d, J = 8.5 Hz, 1H), 7.20 (t, J = 7.5 Hz, 1H), 7.05 (d, J = 12.0 Hz, 2H), 6.75 (d, J = 10.7 Hz, 1H), 5.35 (s, 2H), 3.79 (s, 3H). MS(ESI) (m/z): 350.1(M+H)⁺.

Formula I-25¹H NMR (300 MHz, DMSO) δ 8.41 (d, J = 4.7 Hz, 1H), 8.25 (d, J= 7.9 Hz, 1H), 8.14 (d, J = 7.6 Hz, 1H), 7.65 (t, J = 7.7 Hz, 1H), 7.52 (dd, J = 7.8, 5.0 Hz, 1H), 7.38 (dd, J = 21.0, 7.6 Hz, 3H), 7.20 (dd, J = 16.9, 8.9 Hz, 2H), 5.40 (s, 2H). MS(ESI) (m/z): 338.1(M+H)⁺.。

Example 3: synthesis of Compounds of formulae I-14, I-15, I-16, I-26, I-27

Synthesis of compounds 14d, 15d, 16d, 26d and 27 d:

compound (I)	R
		14e	2-OCH₃
15e	3-OCH₃
		16e	4-OCH₃
26e	2-CH₃; 3-F
		27e	2-CH₃; 5-F

1.0 g (7.24 mmol, 1.0 eq) of 3-fluoro-2-methylbenzaldehyde (compound 26 e) was dissolved in 15 ml of anhydrous ethanol, and 328.6 mg (8.69 mmol, 1.2 eq) of NaBH was added₄After reaction at room temperature for 3.0 hours, the reaction was quenched with a small amount of water, ethanol was distilled off, 15 ml of water was added, extraction was performed with EA (20 ml × 2), washing was performed twice with a saturated sodium chloride solution, anhydrous sodium sulfate was added, stirring and drying were performed, and concentration was performed to obtain 0.7 g of a colorless transparent oil (compound 26 g) with a yield of 69.0%. With reference to the above reaction procedures, 14g, 15g, 16g and 27g of compounds were prepared from 14e, 15e, 16e and 27e, respectively;

0.7 g (4.99 mmol, 1.0 eq) of compound 26g was dissolved in 15 ml CH₂Cl₂1.62 g (5.99 mmol, 1.2 eq) of PBr were added under ice-water bath conditions₃Reacting at room temperature for 1.0 h, and adding a small amount of saturated NaHCO₃The organic phase was washed 1 time, twice with saturated sodium chloride solution, added with anhydrous sodium sulfate, stirred and dried, filtered and evaporated to dryness to give 0.8 g of oil (compound 26 d) in 78.9% yield. The crude product can be directly used for the next reaction without purification. With reference to the above reaction procedures, compounds 14d, 15d, 16d and 27d were prepared from compounds 14g, 15g, 16g and 27g, respectively;

synthesis of Compounds of formulae I-14, I-15, I-16, I-26 and I-27:

formula I	R
		Ⅰ-14	2-OCH₃
Ⅰ-15	3-OCH₃
		Ⅰ-16	4-OCH₃
Ⅰ-26	2-CH₃; 3-F
		Ⅰ-27	2-CH₃; 5-F

Referring to the conditions for the second step of synthesis of compound of formula I-1 in example 1, compounds of formulae I-14, I-15, I-16, I-26 and I-27 are obtained from (oxazolo [5,4-b ] pyridin-2-yl) phenol (compound 1 c) and the corresponding substituted benzyl bromide (compounds 14d, 15d, 16d, 26d and 27 d) in the first step of example 3: (2- ((2-methoxybenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-14); (2- ((3-methoxybenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-15); (2- ((4-methoxybenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-16); (2- ((3-fluoro-2-methylbenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-26); (2- ((5-fluoro-2-methylbenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-27);

formula I-14¹H NMR (400 MHz, DMSO-d ₆) δ 8.35 (dd, J = 5.0, 1.6 Hz, 1H), 8.26 (dd, J = 7.9, 1.6 Hz, 1H), 8.07 (dd, J = 7.8, 1.8 Hz, 1H), 7.69 (dd, J = 7.5, 1.7 Hz, 1H), 7.59 (ddd, J = 8.8, 7.2, 1.8 Hz, 1H), 7.47 (dd, J = 7.8, 5.0 Hz, 1H), 7.28 (dd, J = 13.7, 8.3 Hz, 2H), 7.14 (t, J = 7.7 Hz, 1H), 7.01 (d, J = 8.3 Hz, 1H), 6.98 – 6.92 (m, 1H), 5.26 (s, 2H), 3.79 (s, 3H). MS(ESI) (m/z): 332.1 (M+H)⁺.

Formula I-15¹H NMR (400 MHz, DMSO-d ₆) δ 8.40 – 8.34 (m, 1H), 8.25 (dd, J = 7.8, 1.6 Hz, 1H), 8.08 (dd, J = 7.8, 1.8 Hz, 1H), 7.60 (ddd, J = 8.9, 7.2, 1.8 Hz, 1H), 7.49 (dd, J = 7.9, 4.9 Hz, 1H), 7.38 – 7.07 (m, 5H), 6.88 – 6.80 (m, 1H), 5.32 (s, 2H), 3.75 (s, 3H). MS(ESI) (m/z): 332.1 (M+H)⁺.

Formula I-16¹H NMR (400 MHz, DMSO-d ₆) δ 8.37 (dd, J = 5.0, 1.6 Hz, 1H), 8.26 (dd, J = 7.8, 1.7 Hz, 1H), 8.06 (dd, J = 7.8, 1.9 Hz, 1H), 7.59 (ddd, J= 8.8, 7.3, 1.9 Hz, 1H), 7.52 – 7.44 (m, 3H), 7.35 (d, J = 8.5 Hz, 1H), 7.24 – 7.10 (m, 2H), 6.97 – 6.79 (m, 3H), 5.26 (s, 2H), 3.73 (s, 3H). MS(ESI) (m/ z): 332.1 (M+H)⁺.

Formula I-26¹H NMR (400 MHz, DMSO-d ₆) δ 8.37 (dd, J = 5.0, 1.5 Hz, 1H), 8.26 (dd, J = 7.8, 1.5 Hz, 1H), 8.09 (dd, J = 7.8, 1.7 Hz, 1H), 7.67 – 7.60 (m, 1H), 7.56 (d, J = 7.6 Hz, 1H), 7.51 – 7.42 (m, 2H), 7.28 – 7.09 (m, 3H), 5.34 (s, 2H), 2.27 (d, J = 1.9 Hz, 3H). MS(ESI) (m/z): 334.1 (M+H)⁺.

Formula I-27¹H NMR (300 MHz, DMSO) δ 8.40 (d, J = 4.3 Hz, 1H), 8.24 (d, J= 7.7 Hz, 1H), 8.14 (d, J = 7.4 Hz, 1H), 7.80 – 7.62 (m, 2H), 7.56 – 7.43 (m, 2H), 7.23 (q, J = 8.0 Hz, 2H), 7.05 (t, J = 8.6 Hz, 1H), 5.32 (s, 2H), 2.32 (s, 3H). MS(ESI) (m/z): 334.1 (M+H)⁺.。

Example 4: synthesis of Compounds of formulae I-28, I-29 and I-31

Synthesis of compounds 28d, 29d and 31d;

compound (I)	R
		28f	2-CH₃; 3-OCH₃
29f	2-CH₃; 5-OCH₃
		31f	2,6-dimethyl

1.0 g (6.66 mmol, 1.0 eq) of 2, 6-dimethylbenzoic acid (compound 31 f) was dissolved in 10 ml of dry THF, and 5 ml of solution containing 0.5 g (13.3 mmol, 2.0 eq) of LiAlH (compound 31 f) was added dropwise under ice-bath conditions₄The reaction solution was added dropwise to the dry THF solution and reacted at room temperature for 3.0 hours. Removing solvent, adding 1 mol/L HCl (aq) under ice-water bath condition until no more bubbles are formed, extracting with EA (10 ml. times.2), combining extracts, and adding saturated NaHCO₃Washing for 2 times, washing twice with saturated sodium chloride solution, adding anhydrous sodium sulfate, stirring, drying, concentrating to obtain 0.85 g of oily crude product (compound 31 g), with a yield of 93.7%, directly carrying out the next reaction without purification, and respectively reducing compounds 28f and 29f by referring to the reduction reaction operation to obtain corresponding benzyl alcohol intermediate compounds 28g and 29g;

referring to the conditions for the first step of synthesis of compound 26d in example 3, compounds 28d, 29d and 31d were obtained from compounds 28g, 29g and 31g, respectively, by bromination of phosphorus tribromide, and were used in the next step without purification;

synthesis of compounds of formulae I-28, I-29 and I-31:

formula I	R
		Ⅰ-28	2-CH₃; 3-OCH₃
Ⅰ-29	2-CH₃; 5-OCH₃
		Ⅰ-31	2,6-dimethyl

Referring to the conditions for the second step of the synthesis of compound of formula I-1 in example 1, compounds of formulae I-28, I-29 and I-31 are obtained from (oxazolo [5,4-b ] pyridin-2-yl) phenol (compound 1 c) and the corresponding substituted benzyl bromides obtained in the first step of example 4 (compounds 28d, 29d and 31 d): (2- ((3-methoxy-2-methylbenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-28); (2- ((5-methoxy-2-methylbenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-29); (2- ((2, 6-dimethylbenzyl) oxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-31);

formula I-28¹H NMR (300 MHz, DMSO) δ 8.38 (d, J = 3.9 Hz, 1H), 8.27 (d, J= 7.9 Hz, 1H), 8.08 (d, J = 7.6 Hz, 1H), 7.64 (t, J = 7.8 Hz, 1H), 7.53 – 7.40 (m, 2H), 7.29 (d, J = 7.5 Hz, 1H), 7.18 (t, J = 7.7 Hz, 2H), 6.93 (d, J= 8.0 Hz, 1H), 5.31 (s, 2H), 3.78 (s, 3H), 2.19 (s, 3H). MS(ESI) (m/z): 346.1 (M+H)⁺.

Formula I-29¹H NMR (400 MHz, DMSO-d ₆) δ 8.36 (dd, J = 5.0, 1.6 Hz, 1H), 8.21 (dd, J = 7.8, 1.6 Hz, 1H), 8.09 (dd, J = 7.8, 1.7 Hz, 1H), 7.67 – 7.60 (m, 1H), 7.52 – 7.41 (m, 2H), 7.34 (d, J = 2.7 Hz, 1H), 7.18 (t, J = 7.5 Hz, 1H), 7.09 (d, J = 8.3 Hz, 1H), 6.76 (dd, J = 8.3, 2.7 Hz, 1H), 5.26 (s, 2H), 3.72 (s, 3H), 2.26 (s, 3H). MS(ESI) (m/z): 346.1 (M+H)⁺.

Formula I-31¹H NMR (400 MHz, DMSO-d ₆) δ 8.31 (dd, J = 5.0, 1.7 Hz, 1H), 8.18 (dd, J = 7.7, 1.6 Hz, 1H), 8.03 (dd, J = 7.7, 1.8 Hz, 1H), 7.72 – 7.63 (m, 1H), 7.54 (d, J = 8.5 Hz, 1H), 7.43 (dd, J = 7.8, 5.0 Hz, 1H), 7.16 (dt, J = 25.7, 7.3 Hz, 2H), 7.04 (d, J = 7.5 Hz, 2H), 5.24 (s, 2H), 2.38 (s, 6H). MS(ESI) (m/z): 330.1 (M+H)⁺.。

Example 5: synthesis of Compounds of formula I-30

0.4 g (1.88 mmol, 1.0 eq) of (oxazolo [5,4-b ] pyridin-2-yl) phenol (compound 1 c) was dissolved in 15 mL of acetonitrile, 0.52 g (3.77 mmol, 2.0 eq) of potassium carbonate and 0.57 g (2.26 mmol, 1.2 eq) of 2- (bromomethyl) pyridine hydrobromide (compound 30 d) were added, reaction was carried out at ordinary temperature overnight, the solvent was distilled off, extracting and separating by dichloromethane-water, combining organic phases, washing twice by a saturated sodium chloride solution, adding anhydrous sodium sulfate, stirring and drying, concentrating the filtrate to obtain a crude product, and performing column chromatography by using a PE-EA system column to obtain 0.25 g of white solid 2- (2- (pyridine-2-yl methoxy) phenyl) oxazolo [5,4-b ] pyridine (compound I-30), wherein the yield is 43.7%;

formula I-30¹H NMR (300 MHz, DMSO) δ 8.58 (d, J = 4.6 Hz, 1H), 8.40 (d, J= 3.7 Hz, 1H), 8.31 (d, J = 7.8 Hz, 1H), 8.14 (d, J = 7.7 Hz, 1H), 7.90 (q, J= 7.6 Hz, 2H), 7.63 (t, J = 7.5 Hz, 1H), 7.56 – 7.48 (m, 1H), 7.37 (d, J = 8.2 Hz, 2H), 7.20 (t, J = 7.4 Hz, 1H), 5.41 (s, 2H). MS(ESI) (m/z): 303.1 (M+H)⁺.。

Example 6: synthesis of Compounds of formula I-32

Referring to the first step of the synthesis of compound 1c in the example, 2-aminophenol (compound 32 a) and 2-trifluoromethylphenol (compound 1 b) were used as starting materials to synthesize an intermediate (benzo [ d ] oxazol-2-yl) phenol (compound 32 c);

the compound is 32C m.p.127.2-127.9 ℃ C;¹H NMR (400 MHz, DMSO) δ 11.20 (s, 1H), 8.00 (dd, J = 7.9, 1.6 Hz, 1H), 7.83 (ddd, J = 5.2, 3.0, 1.1 Hz, 2H), 7.53 – 7.41 (m, 3H), 7.13 – 7.04 (m, 2H). MS(ESI) (m/z): 212.1(M+H)⁺ _.

referring again to the second synthesis of formula I-1 in the first step of the example, starting with compound 32c and benzyl bromide (compound 1 d), the (2- (benzyloxy) phenyl) benzo [ d ] oxazole (formula I-32) is obtained;

the formula is I-32 m.p.84.5-86.3 ℃ C;¹H NMR (400 MHz, DMSO) δ 8.05 (dd, J = 7.7, 1.5 Hz, 1H), 7.83 – 7.78 (m, 1H), 7.76 – 7.71 (m, 1H), 7.61 – 7.53 (m, 3H), 7.44 – 7.26 (m, 6H), 7.14 (t, J = 7.5 Hz, 1H), 5.32 (s, 2H). MS(ESI) (m/z): 301.1(M+H)⁺ _。。

example 7: synthesis of Compounds of formula I-33

Referring to the first step of the synthesis of compound 1c in the example, 3-amino-2-hydroxypyridine (compound 1 a) and 4-trifluoromethylphenol (compound 2 b) were used as starting materials to synthesize an intermediate (oxazolo [5,4-b ] pyridin-4-yl) phenol (compound 33 c);

referring again to the second synthesis of formula I-1 in the first step of the example, starting from 33c and benzyl bromide (compound 1 d), the (4- (benzyloxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-33) is obtained;

formula I-33¹H NMR (300 MHz, DMSO) δ 8.33 (d, J = 4.8 Hz, 1H), 8.19 (dd, J= 11.5, 8.6 Hz, 3H), 7.42 (ddd, J = 19.8, 14.1, 6.0 Hz, 6H), 7.26 (d, J = 8.8 Hz, 2H), 5.24 (s, 2H). MS(ESI) (m/z): 302.1(M+H)⁺ _. 。

Example 8: determination of the in vitro inhibition of sphingomyelin synthase 2 by 2-benzyloxyphenyl oxazolopyridines

Experimental apparatus and materials:

1. an electric heating constant temperature water bath (Shanghai-constant technology, Inc.).

2. Vortex mixer (Shanghai Jing Kogyo Co., Ltd. model XW-80A).

3. High speed centrifuge (model Eppendorf 5804R).

4. High performance liquid chromatography Agilent 1100 (Agilent Technologies, Palo Alto, Calif., USA), with quaternary pump, vacuum degassing, FLD fluorescence detector.

5. HPLC column COSMOSIL 5C 18-MS-II (4.6mm I.D. times 250 mm).

6. DMPC, available from Santa Cruz (USA), dissolved in ethanol at a concentration of 40 mmol/L.

7.C6-NBD-Ceramide(6-((N- (7-nitrobenzez-2-oxa-1, 3-diazol-4-yl) amino) hexanoyl) -sphingosine, purchased from Santa Cruz (USA), dissolved in DMSO at a concentration of 1.16 mmol/L.

8. All the organic solvents are purchased from Shanghai national medicine reagent company, the methanol is used as chromatographic pure, the water is filtered by a Milli-Q pump, deionized and ultra-pure water ultrafiltered by a 0.22 mu m membrane, and other biological consumables are purchased from domestic companies.

9. Preparing a solution of a compound to be tested: accurately weighing 1-2 mg of each compound to be detected, and firstly adding a proper amount of DMSO (dimethyl sulfoxide) to accurately prepare a 3 mmol/L stock solution. Taking a certain volume of DMSO stock solution of the compound to be detected, and adding a proper volume of DMSO to dilute the compound to be detected to a solution with a required concentration.

10. SMS1, SMS2 pure enzyme DDM solution and buffer are provided by the national protein science center (Shanghai) Cao Yu subject group.

And (3) detecting the inhibition activity of the 2-benzyloxy phenyl oxazolopyridine compounds on sphingomyelin synthase 2:

mu.L of SMS2 pure enzyme DDM solution (total protein content of 1.5. mu.g/. mu.L), 1. mu.L of DMSO solution or blank DMSO solution of the test compound, 79.7. mu.L of DDM buffer solution were added to a 1.5 mL eppendorf tube, vortexed for 30 seconds, and allowed to stand at room temperature for 5 minutes, followed by 20. mu.L of DDM buffer solution containing 1. mu.L of DMPC in ethanol (40 mmol/L) and 1. mu. L C6-NBD-Ceramide in DMSO solution (1.16 mmol/L), and vortexed for 30 seconds, and then incubated for 0.5 h at 37 ℃ in a water bath. After removal, 200. mu.L of absolute ethanol was added and vortexed for 30 seconds. Taking out 200 μ L of the mixed solution, storing at 4 deg.C, and performing high performance liquid chromatography;

reference (Xiaoodong Deng; Hong Sun; et al.Analytical Letters, 2012, 45 (12)1581-1589.), performing fluorescence quantitative analysis on the prepared sample by adopting a high performance liquid chromatography method the same as the literature, analyzing and recording peak areas Asm value and Acer value of C6-NBD-SM and C6-NBD-Ceramide on corresponding HPLC spectrograms in a blank group sample, a positive control group sample (compound D2) and a compound group sample to be detected, calculating the inhibition rate of the compound to be detected according to the following formula,

the in vitro sphingomyelin synthase 2 inhibitory activity of compounds of formulae I-1 to I-33 was determined according to the above method and the results are:

33) the inhibition rate of (4- (benzyloxy) phenyl) oxazolo [5,4-b ] pyridine (formula I-33) at 10. mu.M is 8%;

half Inhibitory Concentration (IC) of 2-benzyloxyphenyl oxazolopyridine compounds I-1 to I-33 on sphingomyelin synthase 2₅₀) And (3) determination:

gradient dilution is carried out on 3 mmol/L DMSO stock solution of a compound to be tested with the inhibition rate higher than 70% in a single concentration test, 6 concentration gradient solutions are prepared and 1 mu L of the solutions are respectively added into a test system of the first step of the implementation example 6, samples are prepared according to the method of the first step of the implementation example 6, Asm values of the compound under 6 concentrations are measured through a high performance liquid chromatography method, the inhibition rates under 6 concentrations are respectively calculated and fitted to obtain half inhibition concentration IC₅₀Referring to the method for detecting the inhibitory activity of sphingomyelin synthase 2 in the first step of example 8, the test of the single concentration inhibition rate of the compound to be detected is carried out by adopting SMS1 pure enzyme, three groups are tested in parallel, and the average value of the test results is taken;

half Inhibitory Concentration (IC) of the compounds of formulae I-1 to I-33 on sphingomyelin synthase₅₀) And subtype selectivity to SMS1 and SMS2 are shown in Table 1:

TABLE 1 half inhibitory concentration and subtype selectivity of the compounds of formulae I-1 to I-33 for sphingomyelin synthase

Formula I	For SMS2 IC₅₀(μM)	Inhibition rate (100. mu.M) to SMS1
			D2	20.9	57%*¹
Ⅰ-1	16.3	58%*²
			Ⅰ-2	~10	54%*²
Ⅰ-3	2.3**¹	-1%*¹
			Ⅰ-4	>50
Ⅰ-5	>50
			Ⅰ-6	>10
Ⅰ-7	2.3	12%*¹
			Ⅰ-8	>10
Ⅰ-9	1.7	32%*¹
			Ⅰ-10	17.5**¹	55%*²
Ⅰ-11	>10
			Ⅰ-12	~10	1%*²
Ⅰ-13	>100
			Ⅰ-14	19.1**²	62%*²
Ⅰ-15	3.0	1%*¹
			Ⅰ-16	>100
Ⅰ-17	5.0	61%*¹
			Ⅰ-18	~10	59%*²
Ⅰ-19	>50
			Ⅰ-20	>100
Ⅰ-21	>10
			Ⅰ-22	~10	22%*²
Ⅰ-23	>100
			Ⅰ-24	~10
Ⅰ-25	~10
			Ⅰ-26	2.3	11%*¹
Ⅰ-27	~10
			Ⅰ-28	>50
Ⅰ-29	~10
			Ⅰ-30	>100
Ⅰ-31	4.4**¹	53%*¹
			Ⅰ-32	>100
Ⅰ-33	>50

1 SMS1 inhibition rate data N = 3; 2N = 2;

IC50 data N =2 for inhibition activity of 1 SMS 2; 2N = 1;

data without exact values are estimates based on single inhibition rates.

Claims

1. 2-benzyloxy phenyl oxazole pyridine compound shown in formula (I);

in the formula (I), the compound is shown in the specification,

x is selected from nitrogen atom or carbon atom, Y and Z are carbon atom;

r is selected from any one or two of H, halogen, nitro, cyano, methyl, ethyl, methoxy and trifluoromethoxy;

the substituted benzyloxy is in ortho-position or para-position on the benzene ring.

2. The 2-benzyloxyphenyl oxazolopyridine compound represented by the formula (I) according to claim 1, wherein the phenyl group in which R is present is replaced with a pyridyl group.

3. The pharmaceutical composition of the 2-benzyloxy phenyl oxazolopyridine compound with structure of formula (I) as defined in claim 1 or 2 and pharmaceutically acceptable carrier.

4. Use of 2-benzyloxy phenyl oxazolopyridine compound represented by formula (I) as defined in any one of claims 1-2 in preparation of small molecule inhibitor of sphingomyelin synthase.

5. Use of 2-benzyloxyphenyl oxazolopyridine represented by the formula (I) according to any one of claims 1 to 2 in the preparation of a medicament for the prophylaxis or treatment of diseases caused by abnormally increased sphingomyelin levels.

6. The use according to claim 5, wherein said disease caused by an abnormal increase in sphingomyelin levels is atherosclerosis, fatty liver, obesity and type II diabetes and its associated metabolic syndrome.